Pharmaceutical preparations of polypeptides are sensitive to denaturation and degradation upon formulation and storage. Polyols have been used to stabilize proteins and other macromolecules in aqueous formulations and in air drying or lyophilization from aqueous solutions.
U.S. Pat. No. 4,297,344, discloses stabilization of coagulation factors II and VIII, antithrombin III, and plasminogen against heat by adding selected amino acids such as glycine, alanine, hydroxyproline, glutamine, and aminobutyric acid, and a carbohydrate such as a monosaccharide, an oligosaccharide, or a sugar alcohol.
European Patent Application Publication No. 0 303 746 discloses stabilization of growth promoting hormones with polyols consisting of non-reducing sugars, sugar alcohols, sugar acids, pentaerythritol, lactose, water-soluble dextrans, and Ficoll, amino acids, polymers of amino acids having a charged side group at physiological pH, and choline salts.
European Patent Application Publication No. 0 193 917 discloses a biologically active composition for slow release characterized by a water solution of a complex between a protein and a carbohydrate.
Australian Patent Application No. AU-A-30771/89 discloses stabilization of growth hormone using glycine and mannitol.
U.S. Pat. No. 5,096,885 discloses a formulation of hGH for lyophilization containing glycine, mannitol, a non-ionic surfactant, and a buffer.
The use of polyethylene glycols to stabilize proteins is reviewed in Pharm. Res. 8:285-291, 1991.
Examples of the use of trehalose and other polyols for the stabilization of proteins during drying in aqueous systems include the following.
U.S. Pat. No. 4,891,319 discloses the preservation of sensitive proteins and other macromolecules in aqueous systems by drying at ambient temperatures and at atmospheric pressure in the presence of trehalose.
U.S. Pat. No. 5,149,653 discloses a method of preserving live viruses in an aqueous system by drying in a frozen state or at ambient temperature, in the presence of trehalose.
Polyols have also been used to stabilize dry drug formulations as, for example, in WO 8903671, filed May 5, 1989, which discloses the addition of a stabilizer such a gelatin, albumin, dextran, or trehalose to a mixture of a finely powdered drug suspended in a oily medium.
Treatment of a polypeptide with an organic solvent such as methylene chloride poses the problem of denaturation of the polypeptide of interest. Thus, it is an object of this invention to provide a method for stabilizing polypeptides in aqueous formulations treated with organic solvents.
It is another object of the invention to stabilize dry polypeptides treated with organic solvents.
It is another object of the invention to provide a method for stabilization of encapsulated polypeptides.
It is another object of the invention to provide a polypeptide stabilized by an excipient for use in a controlled release formulation, wherein the polypeptide is treated with an organic solvent.